| A cell-free scintillation proximity assay for studies on lysosome-to-phagosome targeting. | |
| | |
MedLine Citation:
|
PMID: 17595222 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Phagocytes, such as macrophages, neutrophils, and dendritic cells, play important roles in the innate immune system through their ability to engulf, kill, and digest invading microbes. In cooperation with the humoral adaptive immune system, coating of substrates with immunoglobulin G (IgG) antibodies enhances several aspects of phagocytosis, including the recognition of substrates by cell surface IgG (Fcgamma) receptors, particle internalization, generation of microbicidal oxygen species, and targeting of lysosomes to phagosomes. We describe a cell-free scintillation proximity assay developed to study the mechanisms of lysosome targeting to phagosomes and the regulation of this process by IgG. The approach involves the use of isolated phagosomes containing scintillant latex beads and lysosomes labeled with a tritiated marker. Scintillation results only when lysosomes and phagosomes come into immediate contact and requires supplementation of reactions with adenosine triphosphate and cytosol; addition of cytosol from IgG-conditioned cells enhances this signal. The method is useful for investigating the biochemistry and regulation of the early tethering and docking steps of lysosome and phagosome interactions. |
| | |
Authors:
|
Vishal Trivedi; Shao Chong Zhang; Walter Stockinger; Axel Nohturfft |
Publication Detail:
|
Type: Journal Article Date: 2007-06-26 |
Journal Detail:
|
Title: Science's STKE : signal transduction knowledge environment Volume: 2007 ISSN: 1525-8882 ISO Abbreviation: Sci. STKE Publication Date: 2007 Jul |
Date Detail:
|
Created Date: 2007-06-27 Completed Date: 2008-01-24 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 100964423 Medline TA: Sci STKE Country: United States |
Other Details:
|
Languages: eng Pagination: pl3 Citation Subset: IM |
Affiliation:
|
Molecular Oncology Research Institute, Tufts-New England Medical Center, Boston, MA 02111, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Adenosine Triphosphate
/
metabolism Cholesterol / analogs & derivatives Cytosol Immunoglobulin G / immunology*, metabolism Lysosomes / metabolism* Microspheres Phagocytosis / immunology* Phagosomes / metabolism* Scintillation Counting / methods Tritium |
| Chemical | |
Reg. No./Substance:
|
0/Immunoglobulin G; 10028-17-8/Tritium; 56-65-5/Adenosine Triphosphate; 57-88-5/Cholesterol; 66106-91-0/cholesteryl oleyl ether |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: GPCRs signaling directly through Src-family kinases.
Next Document: Estrogen regulation of genes important for K+ channel signaling in the arcuate nucleus.